Fig. 6. De novo design of dual-binding site cassettes in U2OS cells.
a 2D density plots (pink-red scale) depicting the predicted Rscore values for one million ML variants binding to (left-to-right): PCP and QCP, MCP and QCP, and MCP and PCP. QCP-PCP dual-binding variants are located in the black dashed square. b Based on the dual-binding mutants for QCP and PCP from our model predictions, we designed an additional cassette. c Results for the dual-binding experiment. Fluorescent foci can be observed for the cassette expressed with either PCP-3xGFP or QCP-3xBFP. For both experiments, both the relevant fluorescent channel and the merged images with the DIC channel are presented. Each of the two experiments were successfully conducted in duplicates and on two different days. Fluorescent wavelengths used in these experiments are: 400 nm for BFP and 490 nm for GFP. d Evaluation of prediction accuracy based on size of the training set. For each training set size, a random set of more than 1,000 training-set variants was withheld for computational testing post-training. Performance is reported as average Pearson correlation over 10 random training and test sets (and standard deviation in shade). Source data are provided as a Source data file.